Intraruminal Injection Research Articles

Efficacy of Alfasure™ for the prevention and treatment of alfalfa bloat in cattle

Alfalfa was grazed at early stages of growth to test the efficacy of field application of the pluronic detergent Alfasure™. Bloat was prevented under optimal weather conditions in the absence of rain but heavy rains could disperse the water-soluble product. A new route of administration of Alfasure™ was also examined for the treatment of bloat. Intraruminal injection of Alfasure™ was completely effective in the treatment of bloat, as was intraruminal injection of an industry standard product but on average the response time of Alfasure™ was significantly faster. Key words: Steers, frothy bloat, Medicago sativa L., feed supplements, Alfasure™

Net flux of folates and vitamin B12 through the gastrointestinal tract of sheep

In Trial 1, three wethers were equipped with ruminal and caecal cannulas, catheters in the right ruminal and caecal veins and a mesenteric artery, as well as blood flow probes around the right ruminal and caecal arteries. Blood samples were collected every 15 min from 30 min before to 60 min after the injection of 9 μmol of folic acid and vitamin B12 per kilogram body weight (BW) in the rumen and after injection of half this dose in the caecum. There was a net release of these vitamins from the rumen shortly after the injection (P < 0.05). In Trial 2, four wethers were equipped with a ruminal cannula, catheters in the right ruminal, mesenteric and portal veins and a mesenteric artery and blood flow probes around the right ruminal and the mesenteric arteries and the portal vein. Blood samples were taken 30 and 60 min before, and 30, 60, 120, 180, 240 and 300 min after the intraruminal injection of 0.32 g of folic acid and 0.98 g of vitamin B12. In Trial 3, blood samples were taken from two wethers previously used in Trial 2, before and 10, 40, 70 and 100 min after the intraruminal injection of 0.32 g of folic acid. These intraruminal injections led to a net release of the vitamins from the mesenteric and portal-drained viscera (P ≤ 0.17) but also from the other gastrointestinal tissues, probably mostly from the duodenum. Therefore, in sheep, the main site of absorption for folic acid is the proximal intestine whereas the absorption of vitamin B12 appears in a more distal part of the small intestine. Key words: Sheep, folic acid, vitamin B12, absorption

The effect of sucrose supplementation on kinetics of nitrogen, ruminal propionate and plasma glucose in sheep

SUMMARYThe effects of dietary sucrose on the metabolic rate of plasma glucose and ruminal propionate as well as the change in nitrogen kinetics were examined in four mature wethers fitted with rumen fistulas in Tsukuba, Japan in 1990. Wethers were fed at 12 equal intervals daily on crushed lucerne hay cubes (1233 g DM/day), with or without 204 g/day of sucrose. Plasma urea and glucose kinetics were determined following a single intravenous injection of [I5N]urea and [U-13C]glucose respectively; and the kinetics of ruminal ammonia and propionate were determined following a single intraruminal injection of [15N]ammonium chloride and [2–13c]sodium propionate respectively. Following supplementation of sucrose to the diet, nitrogen retention was increased (P< 0·05) with a decrease in plasma urea concentration (P< 0·05) and urinary urea excretion (P< 0·05). Sucrose supplementation decreased (P< 005) the concentration and irreversible loss rate of ruminal ammonia. Urinary allantoin excretion did not change with sucrose treatment, but the flow rate of non-ammonia-nitrogen from the rumen was increasedP< 0·05). The transfer rate of ruminal ammonia to plasma urea was also decreased (P< 0·01), whilst the transfer rate of plasma urea to ruminal ammonia was increased (P< 0·05) by dietary sucrose. Sucrose supplementation resulted in a higher concentration of propionate and butyrate (P< 0·05) in the rumen with no significant change in acetate or pH. The concentration of plasma glucose did not change with sucrose treatment, but the concentration of insulin, pool size (P< 0·05) and the irreversible loss rate of glucose (P< 0·01) were increased, reflecting the increase in the production rate of ruminal propionate (P< 0·05). It was concluded that the supplementation of sucrose affected the metabolism of urea and glucose in plasma via a change in ruminal production rate of ammonia and propionate, respectively.

Faeces is a reliable source of body water for measuring tritium in reindeer in summer and in winter

Rates of equilibration and subsequent wash-out of tritium were measured in parallel samples of blood, rumen fluid and faeces collected from two adult female Norwegian reindeer in summer and in winter. The tritium-concentration was the same in all three body water compartments after no more than 9 h following both intravenous and intra-ruminal injection of isotope in summer and following intravenous injection of isotope in winter. The biological half-life of the tritium increased from approximately 3 days in summer to approximately 10 days in winter, probably as a consequence of a decrease in water intake. There were no significant differences in disappearance rates of tritium from blood, rumen fluid and faeces within any of the six experiments. Fresh faeces is therefore a reliable source of body water that can be used in place of blood in studies of body water kinetics in reindeer, thus making it potentially possible to conduct such studies on truly free-living and undisturbed animals.

Weight gain and reproductive performance of spring-born beef heifer calves intraruminally administered oxfendazole1,2

The effect of oxfendazole (Synanthic) on weight gain and reproductive performance of spring-born heifer calves was evaluated at five locations (IL, IN, MI, MO, ND). Three hundred thirty-eight mixed-breed beef heifers (293.4 kg) blocked by weight and age were randomly allotted to one of two treatments. Treatment 1 heifers were dewormed via intraruminal injection of oxfendazole. Treatment 2 heifers served as a control. Heifers were comingled during the winter phase in semi-confinement during the period from late January to late May (winter phase). Heifers were sorted by treatment at the beginning of the grazing season (approximately May 24, 1991) and remained separate until the end of the study (approximately July 31, 1991; summer phase). Dewormed heifers received oxfendazole (4.5 mg/kg BW) in late January and again 28 and 56 d after the beginning of the grazing season. Fecal samples were taken every 21 d from January through September and assayed for gastrointestinal parasite eggs. Fecal egg counts were similar across treatments at the beginning of the trial (P = .34). Deworming decreased fecal egg counts throughout the trial. Twenty-eight days after initial deworming a significant treatment effect on fecal egg counts was observed (P < .01). Winter ADG was significantly higher (P < .01) for dewormed heifers than for controls (.60 vs .52 kg, respectively). Summer ADG was not different between treatments. Dewormed heifers were 7.4 kg heavier than controls (P < .05) at the end of the trial. Age of puberty was not affected by treatment (P = .64). First-service conception and final pregnancy rates were not different between treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Food aversion conditioned in anesthetized sheep

We discovered that a food aversion could be conditioned in anesthetized sheep. Sheep were allowed to eat a familiar food (alfalfa-grain pellets) for 30 min, and 90 min later they were given either an intraruminal (IR) injection of water (C), an IR injection of LiCl (L), anesthesia followed by an IR injection of water (A), or anesthesia followed by an IR injection of LiCl (A+L). Induction of anesthesia was by an intraveneous injection of pentobarbitone sodium, and maintenance of deep anesthesia was by halothane. Sheep were maintained in deep anesthesia for 2 h to ensure that the effects of LiCl on the acquisition of a food aversion, which occur within about 1 h, were completed before they awakened. When tested 5 days later, sheep that received LiCl (treatments L and A+L) consumed less alfalfa-grain pellets than sheep that did not receive LiCl (treatments C and A) (241 g vs. 306 g; p = 0.057). Intake of sheep that were anesthetized (treatments A and A+L) did not differ from that of sheep that were not anesthetized (treatments C and L) (295 g vs. 252 g; p = 0.183). Nor was there an interaction between LiCl and anesthesia ( p = 0.423). Thus, we conclude that changes in preferences for foods caused by postingestive feedback occur automatically every time food is ingested (i.e., they are noncognitive), and the kind and amount of feedback is a function of the match between the food's chemical characteristics and its ability to meet the animal's current demands for nutrients.

Net transfer of urea and ammonia across the ruminal wall of sheep

Texel wethers (68 +/- 2.5 kg BW) fitted with catheters in the ruminal veins and a mesenteric artery, blood flow probes on ruminal arteries, and a ruminal cannula were fed 500 g of orchardgrass hay every 12 h. During the last third of the feeding cycle, intraruminal injections were performed to evaluate the effect of urease activity, osmolality, and concentrations of NH3, butyrate, and CO2 in the rumen on urea and NH3 fluxes across the rumen wall. At pH 6.7, NH3 absorption increased with NH3 and butyrate concentrations in the rumen, and to a lesser extent with CO2 concentration. The increase in ruminal blood flow associated with CO2 and butyrate increase was always greater than the increase in NH3 absorption. Increasing ruminal osmolality slightly decreased NH3 absorption. Ruminal NH3 concentration and ruminal blood flow seemed to be the main determinant of NH3 absorption. Decreasing urease activity in the rumen decreased urea net transfer. The net transfer of urea to the rumen was stimulated by CO2. High concentrations of NH3 (330 mg of N/L) and butyrate (25 mM) in the rumen decreased urea net uptake, whereas osmolality (up to 420 mOsmol/L) did not affect it. Modifications in ruminal blood flow or water net movement across the ruminal wall did not seem to account for the effect of CO2, NH3, and butyrate on urea net uptake.

The effect of deworming naturally infected heifers on liveweight gain, reproductive performance and fecal egg counts

Twenty-six of 52 heifers with an average liveweight of 318 kg were dewormed by intraruminal injection of oxfendazole and placed in drylot on 31 January. The other 26 animals served as controls. All heifers were pastured starting on 14 June and the treatment heifers were dewormed in the same manner as at the start of the experiment (31 January) on 1 July and 31 July. The groups started out with similar total worm eggs per gram of fecal sample, but the dewormed heifers had fewer counts each month thereafter. The drylot average daily gain of the treated heifers was higher (P = 0.01) and less variable (P = 0.12) and by 29 May these animals were heavier by 11.7 kg (P = 0.10). The treated animals did not gain as fast on pasture (P = 0.20), but the gain continued to be less variable (P = 0.09). Age at puberty was not different. Number of open heifers in the treated group was 4 vs. 15% for the control group (P = 0.18). Deworming by intraruminal injection resulted in faster feedlot gains, more uniform gains during both drylot and pasture, reduced parasites and no abscess at injection sites. Key words: Anthelmintics, oxfendazole, cattle, heifers

Fibrinogen-degrading proteins from Haemonchus contortus used to vaccinate sheep

Summary Sixteen nonsibling sheep, approximately 12 months old, that were raised in a helminth-free environment, were used for 2 protection studies 6 months apart. Sheep were vaccinated weekly for 5 weeks by im injection of fibrinogen-degrading proteins derived from the intestinal tract of adult Haemonchus contortus. Ten days after the last vaccination, sheep were given 2,500 infective H contortus larvae by intraruminal injection. Vaccinated sheep produced specific antibodies, and were protected from the worm challenge. Significant differences in mean fecal worm egg counts for 56 days after worm challenge, in mean numbers of H contortus worms, and female fecundity ratios at necropsy were detected in vaccinated sheep, compared with those in control sheep. These data suggest that the fibrinogen-degrading proteins have a protective role in vaccination of sheep against H contortus.

Rate of production of protozoa in the rumen and the flow of protozoal nitrogen to the duodenum in sheep and cattle given a pelleted diet of lucerne hay and barley

SUMMARYTwo crossbred wethers (c. 45 kg) and two Hereford heifers (c. 215 kg) were fitted with permanent cannulas in the rumen and abomasum and given a diet of lucerne hay and barley (60:40). Organic matter (OM) intakes were 0·67 kg/day for sheep and 3·14 kg/day for cattle.The mean numbers of protozoa in the rumen fluid were 9·3 × 105/ml for sheep and 7·4 × 105/ml for cattle. 14C-labelled protozoa were prepared by incubating rumen fluid in vitro with [14C]methyl choline and then isolating them by sedimentation and differential centrifugation. The labelled protozoa were returned to the rumen of each of the donor animals in a single injection. The specific radioactivity in mixed protozoa isolated from the rumen was measured for 3 days. The protozoal pool size was 10 and 12 g N in the sheep and 64 and 44 g N in the cattle. Protozoal N contributed, on average, 48% of the total N in the rumen.Production of protozoal N in the rumen was 9·5 and 11·2 g/day for the sheep and 49·5 and 38·6 g/day for the cattle. The flow of protozoal N from the abomasum was calculated to be 1·6 and 2·9 g/day for sheep and 17·3 and 13·3 g/day for cattle. Thus, on average, 79% of the protozoal N was recycled within the rumen of the sheep and 65% within the rumen of the cattle. Protozoal N made up 11–20% of total N flow from the abomasum. The turnover times of 14C-labelled protozoa injected into the rumen of the sheep and the cattle were similar (26·2 and 26·8 h for sheep; 30·8 and 27·5 h for cattle) and were similar to those for protozoa isolated from the rumen of sheep after direct intraruminal injection of [14C]methyl choline (28 and 36 h).

Induction of tissue eosinophilia by platelet-activating factor in Merino sheep

A study was undertaken on the capacity of platelet-activating factor (PAF) to induce eosinophil accumulation in the mammary glands of non-lactating sheep. Platelet-activating factor induced dose-dependent accumulation of eosinophils in mammary exudates 24 h after infusion. Infection, by intraruminal injection of 20 000 infective Trichostrongylus colubriformis larvae, did not modify the responsiveness of outbred sheep to intramammary infusion of PAF. Mature ewes from high and low responder lines of a flock of sheep, selected on the basis of their responses to vaccination and experimental challenge with T. colubriformis as lambs, did not differ in the magnitude of the eosinophil responses to doses of PAF from 5×10 −13 to 5×10 −7 mol per gland. Intramammary infusion of an extract from third stage larvae of Haemonchus contortus elicited inflammatory exudates containing five- to ten-fold more eosinophils than that elicited by the highest dose of PAF tested. The experiments indicate that the eosinophil chemotactic agonist PAF can induce tissue eosinophilia in sheep and thus may play a role in directing the accumulation of eosinophils in tissues during disease states such as gastrointestinal parasitism.

Methimazole increases the plasma concentrations of the albendazole metabolites of netobimin in sheep.

The influence of methimazole (MTZ) on the pharmacokinetics of netobimin (NTB) and its metabolites was investigated in adult sheep. NTB zwitterion suspension was administered at 20 mg kg-1 by intraruminal injection either alone or with simultaneous administration of MTZ intramuscularly at 1.5 mg kg-1. Blood samples were taken serially over a 120-h period and plasma was analysed by HPLC for NTB, albendazole (ABZ), albendazole sulphoxide (ABZSO), and albendazole sulphone (ABZSO2). NTB parent drug showed fast absorption, low area under the plasma concentration-time curve (AUC) and was rapidly removed from plasma after both treatments. The presence of MTZ did increase significantly the ABZ AUC (138 per cent) and mean residence time (MRT) (86 per cent). Concomitant treatment with MTZ resulted in a notably higher ABZSO plasma profile with significantly longer elimination half-life (t1/2 beta) (390 per cent) and MRT (252 per cent) and with significantly higher AUC (95 per cent). Also, MTZ induced significant increases in ABZSO2 t1/2 beta, AUC, and MRT. We have demonstrated a pharmacokinetic interaction between MTZ and NTB metabolites. MTZ may alter the liver biotransformation of ABZ metabolites which results in pronounced changes in the disposition kinetics of anthelmintically active metabolites.

Protective studies in sheep immunized with cuticular collagen proteins and peptides of Haemonchus contortus

Twenty-eight nonsibling sheep aged approximately 12 months and raised in a helminth-free environment were used in two protection studies. Immunizations were conducted by two intramuscular injections 30 days apart with a synthetic 18AA cuticle collagen peptide and native cuticle collagens derived from the third- and fourth-stage larvae of Haemonchus contortus. Ten days following the last immunization, the sheep were each given 500 infective H. contortus larvae per day for five consecutive days by intraruminal injection. Both collagen materials induced antibodies reactive with cuticle collagens; however, neither induced reproducible protection to H. contortus infections in vaccinated/infected sheep. In the most extensive test, there were no statistical differences in mean faecal worm egg count for 56 days post worm challenge, in mean numbers of H. contortus and female fecundity ratios at necropsy of immunized and unimmunized sheep. Failure to reproducibly immunize sheep with cuticle collagens may be due to the inability of antibodies or host immune cells to reach the collagen epitopes in the nematode cuticle without prior surface coat removal as postulated in human nematode studies.

The toxicity of phomopsin for sheep.

Pure phomopsin was administered to young Merino x Border Leicester wethers by single subcutaneous (SC) and by single and multiple intraruminal (IR) injection. The toxicity after IR injection was influenced by the size of individual doses and the time over which the total dose was given. At high levels of ingestion the toxicity of phomopsin may be limited by absorption rates; with low daily doses the capacity to repair liver damage may be sufficient to prevent cumulative effects. By SC injection a single dose of 10 micrograms/kg approximated the LD50. By IR injection the overall clinical, biochemical and histological responses closest to these of this SC dose resulted from a single dose of 1,000 micrograms/kg. The same total dose administered at daily rates of 50 or 200 micrograms/kg was more toxic and killed all sheep. A single dose of 500 micrograms/kg caused significant liver damage, but no deaths. Single doses of 125 and 250 micrograms/kg and repeated daily doses of 12.5 micrograms/kg over 16 weeks caused no detectable tissue damage. Inappetence was the most sensitive indicator of phomopsin toxicity. About 10% of the sheep differed substantially from the rest of the flock in their susceptibility to phomopsin poisoning.

Resistance of Cooperia curticei against fenbendazole

Resistance of Cooperia curticei against fenbendazole was demonstrated in The Netherlands. Experimental infection of lambs followed by intraruminal injection of 5 mg kg-1 showed a 100 per cent survival of the C curticei population. With an in vitro egg development assay LC50 values between 0.23 and 0.38 micrograms ml-1 thiabendazole were found. This is the first record of a Cooperia species resistant against an anthelmintic.

Survey of anthelmintic resistance in Western Australian sheep flocks. 1. Prevalence.

Between July 1981 and December 1983, 116 randomly selected sheep farms in the south west of Western Australian were surveyed for resistance to anthelmintics. A faecal worm egg count reduction test was carried out on each farm. Anthelmintics tested were thiabendazole (44 mg/kg) and levamisole (7.5 mg/kg) given by intra-ruminal injection and comparisons were made with an untreated group on each farm. Successful tests were carried out on 84 farms and 68% of these had resistant worms present. The prevalence of thiabendazole resistant populations was for H. contortus 18%; Teladorsagia, 41% and Trichostrongylus, 48% and for levamisole resistant populations H. contortus, 10%; Teladorsagia, 41%; Trichostrongylus, 24%, and Nematodirus, 10%. Multiple resistant populations were found on 17% of farms. Although the distribution of nematode genera varied between the 400 to 750 mm and the greater than 750 mm rainfall zones there was no significant difference in the prevalence of resistance between zones. About one third of resistant populations were severely resistant (less than 60% reduction). It is likely that resistant worms were present on many farms without causing clinical disease and continued anthelmintic selection pressure will result in further development of resistance.

The pharmacokinetics of fenbendazole and oxfendazole in cattle.

The pharmacokinetics of fenbendazole and oxfendazole in cattle are described. The pharmacokinetics of oxfendazole were not significantly different when administered orally and by intra-ruminal injection. At a dose rate of 4.5 mg/kg, administered orally, fenbendazole gave rise to mean peak concentrations in plasma of fenbendazole and oxfendazole of 0.11 and 0.13 microgram/ml respectively. Oral administration of oxfendazole, at 4.5 mg/kg body weight, gave rise to plasma peak concentrations of fenbendazole and oxfendazole of 0.10 and 0.20 microgram/ml respectively. Following intra-ruminal administration of oxfendazole, the peak concentrations were 0.11 and 0.18 microgram/ml respectively.

Mebendazole concentrations in sheep plasma

Formulated mebendazole was administered to sheep by intraruminal injection at dose rates of 12.5, 25, 50 or 100 mg/kg bodyweight. The concentrations of mebendazole and two major metabolites were measured by high-performance liquid chromatography in plasma taken at intervals up to 48 hours after treatment. At 12.5 mg/kg the peak plasma concentration was 0.22 +/- 0.03 microM mebendazole, rising to 0.76 +/- 0.04 microM at 100 mg/kg. Peak plasma concentrations occurred between nine and 24 hours for all dose rates and declined rapidly. Two major metabolites were detected; their concentrations exceeded that of mebendazole at all dose rates.

Intraruminal injection of urea and changes in secretion of parotid saliva in sheep.

1. Sheep were injected with varying doses of urea into the rumen and changes in the amount of saliva secreted from one parotid gland were studied. When the dose of urea was 0.1-0.2 g/kg, the secretion underwent no change. When the dose was 0.3 g/kg, the secretion was inhibited. When the dose was 0.4 or 0.5 g/kg, there was a marked inhibition in the secretion. 2. There was a close relationship between the ammonia level of the jugular blood and the inhibition of salivary secretion. When the ammonia level exceeded approximately 0.28 mmol/l, the secretion was inhibited abruptly. 3. Salivary secretion and rumen movement were inhibited when the blood ammonia level exceeded 0.28 mmol/l, after injection of ammonium acetate into the jugular vein. 4. Based on the results mentioned previously, the mechanism of inhibition on the secretion of parotid saliva is discussed in relation to the domestic ruminant in which urea had been injected into the rumen.

Effects of intracarotid or intraruminal injections of NaCl or NaHCO 3 on sodium appetite in goats

Goats, which have been depleted of sodium by loss of parotid saliva from a parotid fistula, have been trained to press panels with their muzzles in order to obatin small volumes of 4% (0.5 M) sodium bicarbonate to drink. Normal goats display no appetite for 4% (0.5 M) bicarbonate solutions. Goats which were depleted of sodium were highly motivated to work for bicarbonate solutions without displaying a marked fall in plasma sodium levels. It was not possible to train sodium depleted goats to press the panels to obtain intracarotid injections of 3% (0.5 M) salt solutions. The continuous intracarotid injection of concentrated 23% (3.9 M) solutions of NaCl, which increased the level of Na + ions in the cephalic circulation by 11 to 18 m-equiv./1., did not reduce the rate at which sodium depleted goats pressed the panel to obtain 4% (0.5 M) bicarbonate solutions to drink. It was not possible to train sodium depleted goats to press the panel switch in order to obtain intraruminal injections of 4% (0.5 M) bicarbonate solution.